{"gene":"WDR64","run_date":"2026-06-11T09:02:06","timeline":{"discoveries":[{"year":2024,"finding":"WDR64, a testis-specific WD40 protein with two β-propellers, interacts with ODF1 to form a protein complex; the WDR64/ODF1 complex localizes at the manchette during nuclear shaping and at the midpiece of mature spermatozoa flagella, implicating WDR64 in manchette and flagellum assembly during spermiogenesis.","method":"Immunoprecipitation–mass spectrometry (IP–MS) to identify interacting partners; immunofluorescence to determine subcellular localization across spermatid developmental steps; RT-PCR and Western blot for expression characterization.","journal":"Heliyon","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — Co-IP/MS interaction plus immunofluorescence localization with developmental context, single lab, no mutagenesis or in vitro reconstitution, no loss-of-function phenotype.","pmids":["39386799"],"is_preprint":false}],"current_model":"WDR64 is a testis-specific WD40 protein that interacts with ODF1 and localizes dynamically from the manchette in elongating spermatids to the midpiece of mature sperm flagella, suggesting a role in manchette-directed intracellular transport and flagellum assembly during spermiogenesis."},"narrative":{"mechanistic_narrative":"WDR64 is a testis-specific WD40-repeat protein with two β-propeller domains that participates in spermiogenesis through a physical association with the outer dense fiber protein ODF1 [PMID:39386799]. The WDR64/ODF1 complex localizes to the manchette during the nuclear-shaping phase of spermatid elongation and subsequently to the midpiece of mature spermatozoan flagella, placing WDR64 at structures that govern manchette-directed intracellular transport and flagellum assembly [PMID:39386799]. Beyond this interaction and its dynamic localization across spermatid development [PMID:39386799], no loss-of-function phenotype, structural model, or biochemical activity has been characterized in the available corpus.","teleology":[{"year":2024,"claim":"Whether the testis-specific protein WDR64 has a defined partner and cellular location during sperm development was unknown; identifying its interactor and localization established a candidate role in manchette and flagellum assembly.","evidence":"IP–MS identification of ODF1 as an interactor plus stage-resolved immunofluorescence, RT-PCR, and Western blot in testis/spermatids","pmids":["39386799"],"confidence":"Medium","gaps":["No loss-of-function or knockout phenotype to establish functional requirement","Interaction shown by Co-IP/MS without reciprocal validation or in vitro reconstitution","Molecular activity of the WD40 β-propellers and the functional consequence of the ODF1 association are undefined"]},{"year":null,"claim":"It remains unknown whether WDR64 is required for manchette-directed transport or flagellum assembly and what cargo or molecular function its β-propellers mediate.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No in vivo gene disruption to test a role in fertility or sperm morphogenesis","No mapping of the WDR64–ODF1 interaction interface or structural characterization"]}],"mechanism_profile":{"molecular_activity":[],"localization":[],"pathway":[],"complexes":[],"partners":["ODF1"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"B1ANS9","full_name":"WD repeat-containing protein 64","aliases":[],"length_aa":1081,"mass_kda":123.6,"function":"","subcellular_location":"","url":"https://www.uniprot.org/uniprotkb/B1ANS9/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/WDR64","classification":"Not Classified","n_dependent_lines":7,"n_total_lines":74,"dependency_fraction":0.0945945945945946},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/WDR64","total_profiled":1310},"omim":[{"mim_id":"621432","title":"WD REPEAT-CONTAINING PROTEIN 64; WDR64","url":"https://www.omim.org/entry/621432"},{"mim_id":"182878","title":"OUTER DENSE FIBER OF SPERM TAILS 1; ODF1","url":"https://www.omim.org/entry/182878"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"","locations":[],"tissue_specificity":"Not detected","tissue_distribution":"Not detected","driving_tissues":[],"url":"https://www.proteinatlas.org/search/WDR64"},"hgnc":{"alias_symbol":["FLJ32978"],"prev_symbol":[]},"alphafold":{"accession":"B1ANS9","domains":[{"cath_id":"-","chopping":"2-96","consensus_level":"high","plddt":72.7459,"start":2,"end":96},{"cath_id":"2.130.10.10","chopping":"132-248_259-380","consensus_level":"medium","plddt":84.8615,"start":132,"end":380}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/B1ANS9","model_url":"https://alphafold.ebi.ac.uk/files/AF-B1ANS9-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-B1ANS9-F1-predicted_aligned_error_v6.png","plddt_mean":70.06},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=WDR64","jax_strain_url":"https://www.jax.org/strain/search?query=WDR64"},"sequence":{"accession":"B1ANS9","fasta_url":"https://rest.uniprot.org/uniprotkb/B1ANS9.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/B1ANS9/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/B1ANS9"}},"corpus_meta":[{"pmid":"39386799","id":"PMC_39386799","title":"WDR64, a testis-specific protein, is involved in the manchette and flagellum formation by interacting with ODF1.","date":"2024","source":"Heliyon","url":"https://pubmed.ncbi.nlm.nih.gov/39386799","citation_count":3,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":2769,"output_tokens":380,"usd":0.007004,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":6612,"output_tokens":822,"usd":0.026805,"stage2_stop_reason":"end_turn"},"total_usd":0.033809,"stage1_batch_id":"msgbatch_01UT1AahFPCAyn2uVweS7A9z","stage2_batch_id":"msgbatch_01HuKWtKNT4Pqj1bc8eEP5RV","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2024,\n      \"finding\": \"WDR64, a testis-specific WD40 protein with two β-propellers, interacts with ODF1 to form a protein complex; the WDR64/ODF1 complex localizes at the manchette during nuclear shaping and at the midpiece of mature spermatozoa flagella, implicating WDR64 in manchette and flagellum assembly during spermiogenesis.\",\n      \"method\": \"Immunoprecipitation–mass spectrometry (IP–MS) to identify interacting partners; immunofluorescence to determine subcellular localization across spermatid developmental steps; RT-PCR and Western blot for expression characterization.\",\n      \"journal\": \"Heliyon\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — Co-IP/MS interaction plus immunofluorescence localization with developmental context, single lab, no mutagenesis or in vitro reconstitution, no loss-of-function phenotype.\",\n      \"pmids\": [\"39386799\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"WDR64 is a testis-specific WD40 protein that interacts with ODF1 and localizes dynamically from the manchette in elongating spermatids to the midpiece of mature sperm flagella, suggesting a role in manchette-directed intracellular transport and flagellum assembly during spermiogenesis.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"WDR64 is a testis-specific WD40-repeat protein with two β-propeller domains that participates in spermiogenesis through a physical association with the outer dense fiber protein ODF1 [#0]. The WDR64/ODF1 complex localizes to the manchette during the nuclear-shaping phase of spermatid elongation and subsequently to the midpiece of mature spermatozoan flagella, placing WDR64 at structures that govern manchette-directed intracellular transport and flagellum assembly [#0]. Beyond this interaction and its dynamic localization across spermatid development [#0], no loss-of-function phenotype, structural model, or biochemical activity has been characterized in the available corpus.\",\n  \"teleology\": [\n    {\n      \"year\": 2024,\n      \"claim\": \"Whether the testis-specific protein WDR64 has a defined partner and cellular location during sperm development was unknown; identifying its interactor and localization established a candidate role in manchette and flagellum assembly.\",\n      \"evidence\": \"IP–MS identification of ODF1 as an interactor plus stage-resolved immunofluorescence, RT-PCR, and Western blot in testis/spermatids\",\n      \"pmids\": [\n        \"39386799\"\n      ],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"No loss-of-function or knockout phenotype to establish functional requirement\",\n        \"Interaction shown by Co-IP/MS without reciprocal validation or in vitro reconstitution\",\n        \"Molecular activity of the WD40 β-propellers and the functional consequence of the ODF1 association are undefined\"\n      ]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"It remains unknown whether WDR64 is required for manchette-directed transport or flagellum assembly and what cargo or molecular function its β-propellers mediate.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"No in vivo gene disruption to test a role in fertility or sperm morphogenesis\",\n        \"No mapping of the WDR64–ODF1 interaction interface or structural characterization\"\n      ]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [],\n    \"localization\": [],\n    \"pathway\": [],\n    \"complexes\": [],\n    \"partners\": [\n      \"ODF1\"\n    ],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"faith_supported":2,"faith_total":2,"faith_pct":100.0}}